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Background: Dermatophytes, fungi of universal distribution, invade semi or fully keratinized structures, such as skin, fur/hair and nails. The various species of dermatophytes are classified into three genera anamorphic: Microsporum, Trichophyton and Epidermophyton. The genus Epidermophyton includes only E. floccosum, that rarely affects animals. The main species responsible for the disease in dogs and cats are Microsporum canis, M. gypseum and Trichophyton mentagrophytes, which were characterized through conventional mycological methodology (microscopic examination with KOH and culture). Molecular methodologies, such as real-time PCR, can contribute to a rapid laboratory diagnosis, helping clinicians to initiate an early antifungal treatment. This case report describes a case of canine dermatophytosis due to Trichophyton mentagrophytes detected from a clinical sample by SYBR-Green real-time PCR.Case: A 8-year-old dog, rescued from the street, was referred to a private veterinary clinic in the city of Canoas, RS, Brazil, presenting generalized lymphadenomegaly, crusted lesions all over the body, generalized alopecia, signs of excoriation and epistaxis. Initially, were administered prednisone [1 mg/kg every 48 h, BID] and cephalexin [30 mg/kg, BID]. Weekly baths with benzoyl peroxide were also given. The therapy was not clinically successful. Wood’s Lamp Test was negative. As a differential diagnosis, PCR for detection of Leishmania was negative. Complete blood count and serum biochemical assay were also performed. For mycological diagnosis, hair specimen was clarified and examined microscopically using 10% potassium hydroxide (KOH) for the visualization of chains of arthroconidia (ectothrix invasion of hair). The infected hair was plated onto MycoselTM Agar, incubated at 28°C for 15 days. Microscopy of hyphae/ conidia and macroscopic colony characteristics (colors and texture) were conducted for the differentiation of the species within the genus Microsporum and Trichophyton. In addition, real-time PCR was applied for direct analysis of the fungal DNA obtained from the hair sample. Microscopic examination was negative. The dermatophyte present in the hair sample was confirmed as Trichophyton mentagrophytes by culture and qPCR (melting-point analysis). The patient was treated with systemic itraconazole [10 mg/kg SID - 90 days]. Twice-weekly application of 2.5 % miconazole and 2% chlorhexidine shampoo until complete cure.Discussion: Dermatophytosis is often listed as self-limiting infection; however, animal dermatophytosis can spread between pets, as well as a zoonotic transmission to humans. The literature on dermatophytosis indicates that Microsporum canis is the predominant etiological agent, followed by M. gypseum. Trichophyon mentagrophytes that appear in a lower percentage of isolation. The culture of hair, even with specific medium containing chloramphenicol and cyclohexamide, may present contaminating fungi, not related to dermatophytosis, which can inhibit or override the growth of dermatophytes. The use of real-time PCR provided a faster and specific diagnosis of dermatophytosis when compared to the conventional mycological methodology for detection and identification of T. mentagrophytes, which takes around 10 to 15 days for culture. It is possible to use this technique as an alternative diagnosis for dermatophytes associated to clinical hair samples of dogs. Keywords: dermatophytosis, dog, pets, qPCR, hair samples, diagnosis, molecular methodology.

Background: Meningoencephalitis in cats is usually related to infectious diseases but may also be caused by the extension of bacterial infections originated in the middle or inner ear. This paper seeks to report on a case of encephalitis due to an infection in the middle/inner ear in a 15-year-old cat. The diagnosis was made through brain CT scan and culture and antibiogram of material collected during a ventral osteotomy of the tympanic bulla and reported grave intravascular haemolysis associated to the treatment with antimicrobial imipenem with cilastatin sodium that has already been described in humans but not in small animals in the researched literature. Case: A 15-year-old, male, castrated, Siamese cat was brought to the Veterinary Clinical Hospital of the UFRGS. The animal presented pyrexia and hyporexia, tested negative in the immunoenzymatic test to detect antigens of the FeLV and antibodies of the FIV. The animal had a history of three episodes of tonic convulsion within two days and otitis that had gone untreated for 10 months prior to the consultation, with apparent spontaneous remission. After blood tests, serum biochemistry profile, abdominal ultrasound scan, chest x-rays and CT scan was requested. It revealed middle and inner otitis in the right ear. It was performed a ventral osteotomy of the right tympanic bulla. Material was collected for culture and antibiogram. It was confirmed that the animal had an inner otitis of bacterial origin by multi-resistant Klebsiella spp. that was sensitive only to antimicrobial imipenem with cilastatin sodium. Fifteen days into the treatment with this antibacterial combination, grave haemolysis was observed, probably associated to the use of the medicine. The patient died on the 17th day of the treatment which was not interrupted as per its owner’s decision. At necropsy, both in the tympanic bulla and liver were found mucous content and whitish lumps. The material collected from the tympanic bulla and cerebrospinal liquid were cultured post-mortem when the bacteria Klebsiella sp. was isolated in the CNS and no bacterial growth was observed in the right tympanic bulla. Discussion: This makes it possible to deduce that there was an intracranial expansion of the otitis which would explain the convulsions and, that due to the use of the antimicrobial agent, the growth of Klebsiella sp. in the tympanic bulla was inhibited. The final diagnosis was bacterial otitis with probable bacterial migration to the brain and liver. Suppurative meningoencephalitis caused by the expansion of the inflammation deriving from middle/inner otitis has been reported as being the cause of convulsions in cats with bacterial isolation. The bacteria that was isolated from the tympanic bulla is in agreement with the microbiota present in middle/inner otitis in cats. Regarding haemolysis, there are no reports that link the use of carbapenems in small animals to it or that mention what the safe therapeutic dosage should be if they were administered. Regarding humans, there have been various reports of haemolytic anaemia caused by this type of medicine including carbapenems. It is also known that, in in vitro tests, imipenem may cause inhibition due to competition from human erythrocyte-glutathione reductase, in charge of defending the erythrocyte against haemolysis. It has been concluded that meningoencephalitis caused by middle/inner otitis must be included as a differential diagnosis in case of convulsions in cats. Additionally, further studies must be carried out in order to determine both the efficacy and the toxicity of carbapenems in veterinary patients, including studies of the occurrence of grave haemolysis after they are administered.

A colibacilose é a principal causa infecciosa de condenação total de carcaça em frangos de corte no sul do Brasil. Esse trabalho tem por objetivo determinar o grau de concordância entre a condenação total por colibacilose de frangos de corte abatidos em estabelecimento sob Serviço de Inspeção Federal (SIF) com o diagnóstico anatomopatológico e bacteriológico. O estudo foi realizado com 45 frangos de corte condenados totalmente por colibacilose (caso) e seus respectivos 45 controles (frangos sem lesões). Em todos os frangos condenados pelo SIF havia lesões macroscópicas e, nos controles não se observou. Através do teste Kappa-Cohen´s essas duas variáveis apresentaram concordância quase perfeita. As aves condenadas apresentaram lesões em fígado (27/45); em fígado e sacos aéreos (11/45); em fígado e coração (2/45); fígado, sacos aéreos e coração (2/45); fígado, sacos aéreos e oviduto (1/45); fígado, sacos aéreos, coração e tecido subcutâneo (1/45); e fígado, sacos aéreos, oviduto e baço (1/45). Observou-se concordância quase perfeita entre condenação e lesão hepática. Histologicamente, em 41 casos e 12 controles observaram- se lesões, sendo os mais frequentes hepatite necrosante aleatória, bronquite fibrino-heterofílica, pericardite aguda e traqueíte linfoplasmocitária. Nas aves com hepatite identificou-se E. coli, Enterococcus sp. e Streptococcus sp. (10/38) e, nas aves com bronquite ou broncopneumonia isolou-se Escherichia coli e Staphylococcus coagulase positiva (9/14). O PCR em tempo real e a imuno-histoquímica para Mycoplasma gallisepticum e M. synoviae foram negativos. Nos casos de condenação total por colibacilose o fígado foi o principal órgão acometido, portanto, o critério de condenação deveria ser revisto, sugerindo condenação por hepatite nesses casos, já que outras bactérias podem causar hepatite, como foi demonstrado nesse estudo. Colibacillosis is the main infectious cause of total carcass condemnation in broilers in southern Brazil. This study aims to determine the degree of agreement between the total carcass condemnation for colibacillosis in broilers slaughtered in establishments under Federal Inspection Service (SIF) with the pathological and bacteriological diagnosis. The study was conducted with 45 broilers totally condemned by colibacillosis (case) and theirs 45 respective controls (chickens without lesions). All broilers condemned had gross lesions and the controls had not. The Kappa-Cohen’s test showed that these two variables had almost perfect agreement. Broilers condemned showed lesions in liver (27/45); liver and air sacs (11/45); liver and heart (2/45); liver, heart and air sacs (2/45); liver, air sacs and oviduct (1/45); liver, air sacs, heart and subcutaneous (1/45); and liver, air sacs, oviduct and spleen (1/45). There is almost perfect agreement between carcass condemnation and liver damage. Histologically, in 41 cases and 12 controls were observed lesions, the most frequent diagnoses were random necrotizing hepatitis, fibrinous-heterophilic bronchitis, acute pericarditis and lymphoplasmacytic tracheitis. In hepatitis cases was isolated Escherichia coli, Enterococcus sp. and Streptococcus sp. (10/38) and in bronchitis or bronchopneumonia E. coli and coagulase positive Staphylococcus (9/14). The polymerse chain reaction (PCR) and immunohistochemistry (IHC) tests for Mycoplasma gallisepticum and M. synoviae were negative. In cases of total carcass condemnation by Colibacillosis the liver was the main organ affected. Therefore, the condemnation criteria should be revised, suggesting conviction for hepatitis in these cases, because other bacteria can cause hepatitis, as demonstrated in this study.

Carcass inspection is important for the detection of certain diseases and for monitoring their prevalence in slaughterhouses. The objective of this study was to assess the occurrence of aspergillosis caused by Aspergillus fumigatus in commercial poultry, through mycological and histopathological diagnosis, and to verify the causal association between the aspergillosis diagnosis criteria and condemnation due to airsacculitis in broilers through a case-control study. The study was carried out with 380 samples. Lungs were collected from broilers that were condemned (95) or not condemned (285) due to airsacculitis directly from the slaughter line. Forty-six (12%) lung samples were positive for A. fumigatus in mycological culture. Among all samples, 177 (46.6%) presented histopathological alterations, with necrotic, fibrinous, heterophilic pneumonia; heterophilic pneumonia and lymphoid hyperplasia being the most frequent. Out of the 380 lungs analyzed, 65.2% (30) showed histopathological alterations and isolation of fungi. The statistical analysis (McNemar's chi-square test) indicated a significant association between the presence of histopathological lesions and the isolation of A. fumigatus. Mycological cultivation and histopathological diagnosis increase the probability of detecting pulmonary alterations in birds condemned by the Final Inspection System, which suggests that such diagnostic criteria can improve the assessment and condemnation of birds affected by airsacculitis. Nos abatedouros, a inspeção das carcaças é fundamental para a detecção e monitoramento da prevalência de certas doenças. Os objetivos do trabalho foram avaliar a ocorrência de aspergilose causada por Aspergillus fumigatus em aves comerciais através do diagnóstico micológico e histopatológico e verificar a possibilidade de associação causal entre os critérios de diagnóstico de aspergilose e condenação por aerossaculite em frangos de corte através de um estudo de caso-controle. O estudo foi realizado com 380 amostras. Foram coletados pulmões de frangos condenados (95) e não condenados (285) por aerossaculite, diretamente na linha de abate de um frigorífico. Quarenta e seis (12%) amostras de pulmão foram positivas na cultura micológica. Do total de amostras, 177 (46,6%) apresentaram alterações histopatológicas, sendo os mais frequentes pneumonia fibrinoheterofílica necrótica, pneumonia heterofílica e hiperplasia linfóide. Do total de 380 pulmões analisados, 65,2% (30) apresentaram alterações histopatológicas e isolamento fúngico. A relação entre a presença de lesões histopatológicas e isolamento de A. fumigatus testada por McNemar indicou que houve associação significativa entre a presença de alterações histopatológicas e o isolamento de A. fumigatus. O cultivo micológico e o exame histopatológico aumentam as chances de se detectar alterações pulmonares em aves condenadas pelo Sistema de Inspeção Final do que nas aves normais, sugerindo que tais critérios de diagnóstico são eficazes para aprimorar a avaliação e condenação de aves por aerossaculite.

Background: Dermatophytoses are cosmopolitan contagious mycoses of the skin and concern a wide range of mammals, including man, and more rarely birds. These mycoses are rarely diagnosed in New World Primates. The most frequent tinea of the subhuman Primates is microsporosis due to Microsporum canis or trichophytosis by Trichophyton mentagrophytes and T. simii. The main clinical features are regular alopecia with erythema and squamosis, usually non-pruriginous although various degree of inammation may modify this typical aspect. As a consequence, an accurate clinical examination, a good differential diagnosis and laboratory analyses are required for a correct identification. Alouatta guariba are primates found from the Amazon region up to the Argentina Norwest. Due to the population development and expansion of the urban perimeters these animals are loosing their space in their own natural habitat and being exposed to more closed relationship with domestic animals and humans. This report contains five cases of dermatophytoses caused by Microsporum canis in Alouatta sp., which were treated in a private clinic in Porto Alegre, RS, Brazil. Case: Five females of Alouatta guariba with aged between 2 and 8 months old were admitted in a private veterinary clinic (Toca dos Bichos, Porto Alegre). Each of them had different injuries (electric chock, death of the parent due gun shot, aggression between families and dog bite) and were admitted in different dates. All five primates were presented with intense pruritus after 7 or 10 days of admission. At physical examination lesions characteristic of dermatophytoses were found. To establish a definitive diagnosis it was collected fur and skin and the material was send to the Mycology Laboratory of the Faculdade de Veterinária, Universidade Federal do Rio Grande do Sul (UFRGS), Porto Alegre, RS, Brazil. The fungic culture was positive for Microsporum canis. The treatment established was fluconazole, 18 mg/kg/day PO, mixed in smashed bananas. After two weeks of treatment, a sensitive clinic improvement was seen in all primates, characterized by diminishing of pruritus and alopecic areas. At 30 days of treatment the animals had no clinical signs of lesions and had their fur completely re-grown. The medication was administered for more 15 days, totalizing 45 days of treatment, at which time the animals were considered cured. The primates were monitored for more 30 days after the last dose of fluconazole. Discussion: Case reports on the isolation of Microsporum canis in non-human primates, mainly in New World Primates, are very rare in the Brazilian literature. It is necessary more cohesive approach to nonhuman primate (NHP) dermatology, without relying on assumptions that it is similar to other veterinary disease. Mycological culture remains the gold standard for the diagnosis of animal dermatophytosis and the only method for the phenotypic identication of dermatophyte species. The fluconazole treatment proved to be highly effective, as the animals were all cured and there was no side effects related. This case report proves the importance of a correct mycological diagnostic in free-range animals as an effective treatment in Alouatta guariba.

Pneumocystis has been isolated from a wide range of unrelated mammalian hosts, including humans, domestic and wild animals. It has been demonstrated that the genome of Pneumocystis of one host differs markedly from that of other hosts. Also, variation in the chromosome and DNA sequence of Pneumocystis within a single host species has been observed. Since information about the occurrence and nature of infections in wild animals is still limited, the objective of this work was to detect the presence of Pneumocystis sp. in lungs of bats from two states from Brazil by Nested-PCR amplification. The bats, captured in caves and in urban areas, were obtained from the Program of Rabies Control of two States in Brazil, Mato Grosso and Rio Grande do Sul, located in the Mid-Western and Southern regions of the country, respectively. DNAs were extracted from 102 lung tissues and screened for Pneumocystis by nested PCR at the mtLSU rRNA gene and small subunit of mitochondrial ribosomal RNA (mtSSU rRNA). Gene amplification was performed using the mtLSU rRNA, the primer set pAZ102H - pAZ102E and pAZ102X - pAZY, and the mtSSU rRNA primer set pAZ102 10FRI - pAZ102 10R-RI and pAZ102 13RI - pAZ102 14RI. The most frequent bats were Tadarida brasiliensis (25), Desmodus rotundus (20), and Nyctinomops laticaudatus (19). Pneumocystis was more prevalent in the species Nyctinomops laticaudatus (26.3% = 5/19), Tadarida brasiliensis (24% = 6/25), and Desmodus rotundus (20% = 4/20). Besides these species, Pneumocystis also was detected in lungs from Molossus molossus (1/11, 9.1%), Artibeus fimbriatus (1/1, 100%), Sturnira lilium (1/3, 33.3%), Myotis levis (2/3, 66.7%)and Diphylla ecaudata (1/2, 50%). PCR products which could indicate the presence of Pneumocystis (21.56%) were identified in DNA samples obtained from 8 out of 16 classified species from both states (5 bats were not identified). This is the first report of detection of Pneumocystis in bats from Brazil. Pneumocystis tem sido isolado de uma grande variedade de hospedeiros mamíferos, incluindo humanos, animais domésticos e selvagens. Tem se demonstrado que o genoma do Pneumocystis de um hospedeiro difere marcadamente do de outros, assim como há variação no cromossomo e na seqüência de DNA dentro de uma única espécie de hospedeiro. Sabendo que a informação da ocorrência e natureza da infecção em animais silvestres ainda é limitada, o objetivo do trabalho foi detectar, por Nested-PCR, a presença de Pneumocystis sp. em pulmões de diferentes espécies de morcegos de dois estados do Brasil. Estes mamíferos voadores foram capturados em cavernas, áreas florestadas, de campo e urbanas pelo Programa de Controle da Raiva do Mato Grosso (região Centro-Oeste) e do Instituto de Pesquisas Veterinárias Desidério Finamor (RS) e Instituto Sauver no Rio Grande do Sul (região Sul). Os DNAs foram extraídos de 102 pulmões e realizado Nested-PCR utilizando os primers pAZ102H-pAZ102E e pAZ102X/R1-pAZY/R1 para amplificação do gene mtLSU-rRNA, e pAZ102 10F-RI - pAZ102 10R-RI e pAZ102 13-RI - pAZ14-RI para amplificação do gene mtSSU-rRNA. As espécies mais freqüentes foram Tadarida brasiliensis (25), Desmodus rotundus (20) e Nyctinomops laticaudatus (19). Pneumocystis foi detectado com maior prevalência nas Nyctinomops laticaudatus (26,3% = 5/19), Tadarida brasiliensis (24% = 6/25) e Desmodus rotundus (20% = 4/20). Além destas espécies, Pneumocystis foi também detectado nos pulmões de Molossus molossus (1/11, 9,1%), Artibeus fimbriatus (1/1, 100%), Sturnira lilium (1/3, 33%), Myotis levis (2/3, 66,7%)e Diphylla ecaudata (1/2, 50%). Os produtos de PCR indicaram a presença de Pneumocystis (21.56%) em amostras obtidas de 8 das 16 espécies classificadas para ambos os estados (cinco morcegos não foram classificados). Este é o primeiro registro de detecção de Pneumocystis em morcegos no Brasil.

Descrevem-se dois surtos de uma doença neurológica que afetou rebanhos bovinos leiteiros que consumiam bagaço de malte contaminado por Aspergillusclavatus no município de Viamão, estado do Rio Grande do Sul. A morbidade em ambos os surtos foi em torno de 30% e a letalidade, 50% e 100%. A evolução clínica da doença variou de 5 a 64 dias. Dentre os animais que se recuperaram apenas um permaneceu com seqüelas locomotoras leves. Os sinais clínicos eram predominantemente locomotores e incluíam tremores musculares de intensidade variável, hiperestesia e membros pélvicos com ataxia, paresia e paralisia progessivas, e apoio sobre os boletos. Os distúrbios locomotores eram intensificados pelo exercício que, em geral, desencadeava quedas. Havia também marcada queda na produção leiteira, no entanto o apetite e a dipsia eram mantidos até próximo da morte ou eutanásia. Cinco bovinos foram necropsiados e destes dois apresentaram lesões macroscópicas nos músculos esqueléticos, principalmente nos membros pélvicos e torácicos caracterizadas por alterações necróticas e mineralização. No sistema nervoso, os principais achados consistiam de degeneração e necrose neuronal cromatolítica em núcleos nervosos específicos no tronco encefálico, nos cornos ventrais da medula espinhal e nos gânglios trigeminal, estrelado, celíaco e espinhais. Em dois bovinos havia adicionalmente degeneração walleriana nos funículos dorsais da medula espinhal e nervos isquiádico e fibular. O diagnóstico foi baseado nos dados epidemiológicos, sinais clínicos, achados de necropsia, histopatológicos e micológicos. Os aspectos epidemiológicos, clínicos e patológicos da enfermidade, além de possíveis mecanismos patogenéticos e diagnósticos diferenciais são discutidos. Two outbreaks of a neurological disease affecting herds of dairy cattle that were fed moldy beer residues contaminated with Aspergillus clavatus in the county of Viamão, Rio Grande do Sul, Brazil, are described. The morbidity of both outbreaks was 30% and the lethality 50% and 100%. The clinical course varied from 5 to 64 days. Only one of the animals that recovered from the disease remained with slight locomotor sequels. Clinical signs were predominantly locomotor and included muscle tremors of varied intensity, hyperesthesia and progressive posterior ataxia, paresis and paralysis with knuckling of fetlocks of the hind limbs. Gait abnormalities were more pronounced after exercises which in general led to falling down. There was also reduced milk production, but appetite and water intake were maintained until close to death or euthanasia. From five cattle necropsied, two showed macroscopic lesions characterized by necrotic changes and mineralization in pelvic muscles and thoracic limbs. The main histological findings consisted of chromatolytic neuronal degeneration and necrosis in selected nuclei of the brain stem, the ventral horn of the spinal cord, and of the trigeminal, stellate celiac and spinal ganglions. In two cattle there was wallerian degeneration in dorsal funiculi of the spinal cord and ischiadic and fibular nerves. The diagnosis was based on epidemiological data, clinical signs, necropsy findings, histological lesions and mycological examination. Epidemiologic, clinical and pathologic aspects, pathogenetic mechanisms and differential diagnoses are discussed.

Pneumocystis tem sido isolado de uma grande variedade de hospedeiros mamíferos, incluindo humanos, animais domésticos e selvagens. Tem se demonstrado que o genoma do Pneumocystis de um hospedeiro difere marcadamente do de outros, assim como há variação no cromossomo e na seqüência de DNA dentro de uma única espécie de hospedeiro. Sabendo que a informação da ocorrência e natureza da infecção em animais silvestres ainda é limitada, o objetivo do trabalho foi detectar, por Nested-PCR, a presença de Pneumocystis sp. em pulmões de diferentes espécies de morcegos de dois estados do Brasil. Estes mamíferos voadores foram capturados em cavernas, áreas florestadas, de campo e urbanas pelo Programa de Controle da Raiva do Mato Grosso (região Centro-Oeste) e do Instituto de Pesquisas Veterinárias Desidério Finamor (RS) e Instituto Sauver no Rio Grande do Sul (região Sul). Os DNAs foram extraídos de 102 pulmões e realizado Nested-PCR utilizando os primers pAZ102H-pAZ102E e pAZ102X/R1- pAZY/R1 para amplificação do gene mtLSU-rRNA, e pAZ102 10F-RI - pAZ102 10R-RI e pAZ102 13-RI - pAZ14- RI para amplificação do gene mtSSU-rRNA. As espécies mais freqüentes foram Tadarida brasiliensis (25), Desmodus rotundus (20) e Nyctinomops laticaudatus (19). Pneumocystis foi detectado com maior prevalência nas Nyctinomops laticaudatus (26,3% = 5/19), Tadarida brasiliensis (24% = 6/25) e Desmodus rotundus (20% = 4/20). Além destas espécies, Pneumocystis foi também detectado nos pulmões de Molossus molossus (1/11, 9,1%), Artibeus fimbriatus (1/ 1, 100%), Sturnira lilium (1/3, 33%), Myotis levis (2/3, 66,7%) e Diphylla ecaudata (1/2, 50%). Os produtos de PCR indicaram a presença de Pneumocystis (21.56%) em amostras obtidas de 8 das 16 espécies classificadas para ambos os estados (cinco morcegos não foram classificados). Este é o primeiro registro de detecção de Pneumocystis em morcegos no Brasil. Pneumocystis has been isolated from a wide range of unrelated mammalian hosts, including humans, domestic and wild animals. It has been demonstrated that the genome of Pneumocystis of one host differs markedly from that of other hosts. Also, variation in the chromosome and DNA sequence of Pneumocystis within a single host species has been observed. Since information about the occurrence and nature of infections in wild animals is still limited, the objective of this work was to detect the presence of Pneumocystis sp. in lungs of bats from two states from Brazil by Nested-PCR amplification. The bats, captured in caves and in urban areas, were obtained from the Program of Rabies Control of two States in Brazil, Mato Grosso and Rio Grande do Sul, located in the Mid-Western and Southern regions of the country, respectively. DNAs were extracted from 102 lung tissues and screened for Pneumocystis by nested PCR at the mtLSU rRNA gene and small subunit of mitochondrial ribosomal RNA (mtSSU rRNA). Gene amplification was performed using the mtLSU rRNA, the primer set pAZ102H - pAZ102E and pAZ102X - pAZY, and the mtSSU rRNA primer set pAZ102 10FRI - pAZ102 10R-RI and pAZ102 13RI - pAZ102 14RI. The most frequent bats were Tadarida brasiliensis (25), Desmodus rotundus (20), and Nyctinomops laticaudatus(19). Pneumocystis was more prevalent in the species Nyctinomops laticaudatus (26.3%=5/19),Tadarida brasiliensis (24% = 6/25), and Desmodus rotundus (20% = 4/20). Besides these species, Pneumocystis also was detected in lungs from Molossus molossus(1/11 9.1%), Artibeus fimbriatus (1/1, 100%), Sturnira lilium (1/3, 33.3%), Myotis levis(2/3, 66.7%) and Diphylla ecaudata (1/2,50%). PCR products which could indicate the presence of Pneumocystis(21.56%) were identified in DNA samples obtained from 8 out of 16 classified species from both states(5 bats were not identified). This is the first report of detection of Pneumocy in bats from Brazil.

Os dermatófitos são responsáveis pela doença fúngica mais frequente na clínica médica veterinária de pequenos animais: a dermatofitose. Trata-se de uma zoonose de distribuição mundial, caracterizada pela infecção de estruturas queratinizadas, como pele, cabelos, pelame, unhas e garras. O cultivo micológico é o padrão de referência para o diagnóstico dessa micose, no entanto requer aproximadamente 15 dias para um resultado definitivo. Métodos de diagnóstico moleculares baseados em PCR possuem elevada sensibilidade e especificidade, além de proporcionarem resultados entre um a três dias. O objetivo deste estudo foi realizar a detecção molecular de dermatófitos diretamente de amostras de pelame de cães e de gatos com suspeita de dermatofitose e comparar os resultados obtidos com o método padrão de referência. Foram coletadas 219 amostras de pelame, as quais foram submetidas ao cultivo micológico e à extração de DNA para realização de análise por qPCR. A comparação entre testes mostrou concordância de resultados em 70% das amostras: a qPCR identificou corretamente amostras positivas em cultivo em 34% dos casos, enquanto identificou corretamente amostras negativas em cultivo em 95% dos casos. Além disso, a qPCR foi capaz de detectar dermatófitos em sete amostras negativas em cultivo. A qPCR detectou maior proporção de dermatófitos em amostras de gatos (20%) do que em amostras de cães (13%). Diferentes métodos prévios à extração de DNA (tratamento por congelamento versus sem tratamento) e uso de amostras com diferentes períodos entre coleta e extração de DNA (menor ou igual a um ano versus maior que um ano) foram comparados, observando-se que o uso de amostras frescas associado ao tratamento por congelamento prévio à extração de DNA proporcionou 64% de positividade na qPCR das amostras positivas em cultivo. Ainda, amostras de alta pureza e concentração de DNA apresentaram 82% de positividade na qPCR das amostras positivas em cultivo. A qPCR pode ser utilizada como um teste de triagem inicial prévio ao cultivo micológico. Ainda assim, a técnica molecular desenvolvida deve ser adaptada com a adição de novos métodos juntos à extração a fim de aumentar sua eficácia, podendo se tornar o método de escolha para a detecção de dermatófitos em amostras clínicas no futuro. Dermatophytes are responsible for the most common fungal disease in small animal veterinary practice: dermatophytosis. It is a worldwide zoonosis characterized by infection of keratinized structures such as skin, hair, fur, nails and claws. Mycological culture is the reference standard for diagnosis of this ringworm, however it requires at least 15 days for a definitive result. PCR-based diagnostic methods are high sensitivity and specificity and provide results within one to three days. The aim of this study was to perform molecular detection of dermatophytes directly from dog and cat fur samples with suspected dermatophytosis and to compare results with reference standard method. A total of 219 fur samples were collected and submitted to mycological culture and DNA extraction for qPCR analysis. Comparison between tests showed results agreement in 70% of samples: qPCR correctly identified positive samples in culture in 34% of cases; while correctly identified negative culture samples in 95% of cases. In addition, qPCR was able to detect dermatophytes in seven negative culture samples. qPCR detected a higher proportion of dermatophytes in cat (20%) than in dog (13%) samples. Different methods prior to DNA extraction (freezing treatment versus untreated) and use of samples with different periods between DNA collection and extraction (less than or equal to one year versus more than one year) were compared: use of fresh samples associated with freezing treatment prior to DNA extraction yielded 64% positivity in qPCR of positive samples in culture. Also, high purity and high amount of DNA samples yielded 82% positivity in qPCR of positive samples in culture. qPCR can be used as initial screening test prior to mycological culture. Nevertheless, this molecular technique developed must be adapted with addition of new methods together with extraction in order to increase its effectiveness and could become the method of choice for dermatophytes detection in clinical samples in future.